Blubber Samples Research Articles

Space-use strategy drives fine-scale spatial variation of chlorinated paraffins in indo-pacific humpback dolphins

Contaminant accumulation in organisms can be influenced by both biological traits and environmental conditions. However, delineating the main factors affecting contaminant burdens in organisms remains challenging. Here, we conducted an initial investigation into the impact of diet and habitat on the accumulation of short- (SCCPs) and medium-chain chlorinated paraffins (MCCPs) in Indo-Pacific humpback dolphins (2003–2020, n = 128) from the Pearl River Estuary (PRE), a highly polluted estuary in China. The detected levels of SCCPs (5897 ± 3480 ng g−1 lw) and MCCPs (13,960 ± 8285 ng g−1 lw) in blubber samples of humpback dolphin are the highest among recorded values marine mammals. Both SCCPs and MCCPs exhibited biomagnification factor values exceeding 1, suggesting their biomagnification potential within the dolphins and their diet. Quantitative diet analysis using the dolphin fatty acid signatures revealed that humpback dolphins inhabiting the western PRE consumed a larger proportion of carnivorous fish than those from the eastern PRE. However, spatial analysis showed that humpback dolphins in the western PRE contained lower SCCP/MCCP concentrations than those from the eastern PRE. Based on these findings we suggest that, compared to diet differences, spatial variations of SCCPs/MCCPs in humpback dolphins may be predominantly influenced by their space-use strategies, as the eastern PRE is closer to the pollutant discharge source and transfer routes.

Detection of phthalate esters and targeted metabolome analysis in Franciscana dolphin (Pontoporia blainvillei) blubber in the coast of Santa Catarina, southern Brazil

The concerning of plastic pollution in different ecosystems has been worsened by the widespread presence. Phthalate esters (PAEs), plasticizers found in everyday products, can migrate into the environment, especially into the oceans. Researches on their effects on cetaceans are still rare. Metabolomics helps assess perturbations induced by exposure to PAEs, which act as persistent endocrine disruptors. Four PAEs (dimethyl phthalate - DMP, diethyl phthalate - DEP, dibutyl phthalate - DBP, and di(2-ethylhexyl phthalate – DEHP) were analyzed, along with cholesterol and fatty acid profiles of P. blainvillei's blubber samples collected in southern Brazil. The study reveals pervasive contamination by PAEs - especially DEHP, present in all samples - with positive correlations between DEP content and animal size and weight, as well as between the DEHP amount and the C17:1 fatty acid. These findings will be relevant to conservation efforts aimed at this threatened species and overall marine ecosystems.

High Amounts of Halogenated Natural Products in Sperm Whales (Physeter macrocephalus) from Two Italian Regions in the Mediterranean Sea.

Halogenated natural products (HNPs) are considered to be emerging contaminants whose environmental distribution and fate are only incompletely known. Therefore, several persistent and bioaccumulative HNP groups, together with man-made polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs), were quantified in the blubber of nine sperm whales (Physeter macrocephalus) stranded on the coast of the Mediterranean Sea in Italy. The naturally occurring polybrominated hexahydroxanthene derivatives (PBHDs; sum of TetraBHD and TriBHD) were the most prominent substance class with up to 77,000 ng/g blubber. The mean PBHD content (35,800 ng/g blubber) even exceeded the one of PCBs (28,400 ng/g blubber), although the region is known to be highly contaminated with man-made contaminants. Based on mean values, Q1 ∼ PBDEs > MeO-BDEs ∼ 2,2'-diMeO-BB 80 and several other HNPs followed with decreasing amounts. All blubber samples contained an abundant compound whose molecular formula (C16H19Br3O2) was verified using high-resolution mass spectrometry. The only plausible matching isomer was (2S,4'S,9R,9'S)-2,7-dibromo-4'-bromomethyl-1,1-dimethyl-2,3,4,4',9,9'-9,9'-hexahydro-1H-xanthen-9-ol (OH-TriBHD), a hydroxylated secondary metabolite previously detected together with TriBHD and TetraBHD in a sponge known to be a natural producer of PBHDs. The estimated mean amount of the presumed OH-TriBHD was 3000 ng/g blubber, which is unexpectedly high for hydroxylated compounds in the lipids of marine mammals.

Oil Identification of harp seal and other select marine mammals

Because of the rich omega-3 fatty acids content, harp seal (Pagophilus groenlandicus) oil is a popular supplement that is packaged as pills in Canada and sold for medicinal purposes, although this practice is banned in the United States. Due to US regulations, it is important to be able to distinguish between fish oil and seal oil, but the taxonomic determination of oils provenance has been a difficult problem to solve. In this study, Direct Analysis in Real Time time-of-flight mass spectrometry (DART TOFMS) was used to analyze the chemotypes of blubber samples collected from seven species of marine mammals, including seals, sea lions, and a porpoise. Results indicated that the chemotype profiles found in negative-ion mode could be used to separate all of the species using Discriminant Analysis of Principal Components (DAPC). Consequently, this study suggests that it may be possible to identify the taxonomic source of marine mammal oils based on chemical chemotypes.

Monk seals (Monachus monachus) in the Mediterranean Sea: The threat of organochlorine contaminants and polycyclic aromatic hydrocarbons

The Mediterranean monk seal (Monachus monachus) is an emblematic species of conservation concern. Anthropogenic pressures have led to severe population declines and significant fragmentation of the remaining populations. Because of their close relationship with coastal areas, Mediterranean monk seals may be potentially exposed to pollution from agricultural sources, as well as from oil tanker spills. Although monitoring of pollution has long been considered a priority for this species, data on monk seal contamination levels are scarce. In this study, 55 blubber samples of all genders and age classes collected during necropsies (1995–2013) from seals of the Eastern Mediterranean subpopulation were analyzed for organochlorine compounds (OCs), i.e., hexachlorobenzene (HCB), polychlorinated biphenyls (PCBs) and dichlorodiphenyltrichloroethane (DDTs), and polycyclic aromatic hydrocarbons (PAHs). Overall, PCBs > DDTs ≫ PAHs > HCB in all samples. Results showed a significant downward trend over the 19-year period for DDTs and HCB. No marked pattern was found for PAHs, even though relative abundance of cancerogenic PAH fraction rose in recent years. PCB levels in subadult specimens increased noticeably over time despite worldwide ban. Our findings did not suggest recent releases of DDT or new pesticides (e.g., Dicofol) in the environment, but may indicate an improper disposal of ancient storages of PCBs or a remobilization from reservoirs. OC levels exceeded thresholds that are commonly associated with immunosuppression and reproductive impairment in other pinnipeds. Hence, OCs could be responsible of potential toxicological effects in this subpopulation. This study represents the first report on PAH and one of the few reports on OC bioaccumulation in Mediterranean monk seals. Based on the levels found in the blubber of monk seals from the Eastern Mediterranean, and given the known harmful effects associated to the prolonged exposure to these substances and the reduced long-term expectations of species recovery, regular monitoring is advocated.

Can blubber steroid hormone measurements reveal reproductive state in narwhals?

Abstract Hormone measurements from blubber samples have been used to assess reproduction in cetaceans and are a widely applicable technique, as blubber samples are often collected from necropsied individuals and biopsies are readily collected from live, free-swimming cetaceans. Many studies have assessed reproduction in cetaceans based on blubber hormone concentrations but few have validated their findings with individuals of known reproductive state. The objectives of this study were to use a unique dataset of paired female narwhal (Monodon monoceros) reproductive tracts and blubber samples to: (1) determine narwhal reproductive state based on ovarian analysis; (2) evaluate progesterone, estradiol, testosterone and corticosterone concentrations in paired blubber samples to validate the use of blubber hormone measurements as a technique to assess reproductive state in narwhals; and (3) determine narwhal reproductive rates using reproductive tract and hormone analyses. Female narwhals with complete reproductive tracts or known ages (n = 13) were categorized as: pregnant (fetus or placenta present; n = 5), active (at least one corpus luteum present; n = 2), resting (at least one corpus albicans present; n = 3) or immature (absence of corpora lutea and albicantia or age <8; n = 3), and eight individuals were classified as unknown due to incomplete reproductive tracts. Estradiol, testosterone, and corticosterone concentrations were not useful for assessing reproductive state; however, progesterone concentrations were higher in pregnant narwhals (432.66 ± 182.13 ng/g) than active (1.57 ± 0.42 ng/g), resting (1.52 ± 0.87 ng/g) and immature (1.44 ± 0.71 ng/g) individuals, validating the use of blubber progesterone concentrations in determining pregnancy in narwhals. Using a progesterone threshold for pregnancy, determined in this study, we were able to classify three individuals with incomplete reproductive tracts as pregnant and identify a potential impending pregnancy loss. The results from this study suggest that blubber progesterone concentrations are useful for assessing pregnancy and can help inform reproductive rates of narwhal populations.

Optimization and validation of a micro–QuEChERS method for phthalates detection in small samples of cetacean blubber

In this study, an innovative method was developed to detect and quantify phthalates in fresh cetacean blubber. An adaptation of the ammonium formate QuEChERS method was used and adapted as a micro-extraction for small quantities of samples. Significantly, this technique utilized minimal quantities of reagents and salts, with the additional implementation of rigorous Quality Assurance/Quality Control protocols to further reduce background contamination. To ensure the reliability of this method, comprehensive validation procedures were conducted, with a specific focus on two widely studied cetacean species: the common bottlenose dolphin (Tursiops truncatus) and the short-finned pilot whale (Globicephala macrorhynchus). Determination coefficients (R2) for matrix-matched calibration were >0.93 with limits of quantifications (LOQ) of the method in the range of 5–10 ng/g. Mean recovery values were between 40 and 100 %. This novel methodology holds particular relevance for environmental research studies, offering the capability to detect emerging contaminants with minimal sample requirements. This aspect is particularly valuable in investigations that involve free-ranging animals and rely on biopsy sampling. It allows for the assessment of contaminant levels in healthy individuals within wild populations, enhancing our understanding of ecological impacts and potential conservation measures.•A micro-extraction adaptation of the ammonium formate QuEChERS method was developed and applied to a small quantity of fresh cetacean blubber to detect phthalates.•Small quantities of reagents and salts were used, and additional Quality Assurance/ Quality Control procedures were taken to further minimize background contamination.•Method validation was carried out for two cosmopolitan and extensively studied cetacean species: the common bottlenose dolphin (Tursiops truncatus) and the short-finned pilot whale (Globicephala macrorhynchus).

Pharmaceutical residues in stranded dolphins in the Bay of Biscay

There is a growing concern about the presence of pharmaceuticals on the aquatic environment, while the marine environment has been much less investigated than in freshwater. Marine mammals are suitable sentinel species of the marine environment because they often feed at high trophic levels, have unique fat stores and long lifespan. Some small delphinids in particular serve as excellent sentinel species for contamination in the marine environment worldwide. To the best of our knowledge, no pharmaceuticals have been detected or reported in dolphins so far.In the present study, muscle, liver and blubber samples from three common dolphins (Delphinus delphis) and seven striped dolphins (Stenella coeruleoalba) stranded along the Basque Coast (northern Spain) were collected. A total of 95 pharmaceuticals based on detectability and predicted ability to bioaccumulate in fish were included in the liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis.At least one pharmaceutical was found in 70 % of the individuals. Only three of the 95 monitored pharmaceuticals were detected in dolphin's tissues. Very low concentrations (<1 ng/g) of orphenadrine and pizotifen were found in liver and promethazine in blubber.Herein, the gap in the knowledge regarding the study organisms and marine environments with respect to pharmaceutical pollution, which demands further research to understand if pharmaceuticals are a threat for these apex predators, is highlighted and discussed.

Mobilization of persistent organic pollutants in humpback whales: Insights from feeding areas in the Antarctic peninsula and Strait of Magellan to migration, breeding, and calving grounds along the Brazilian coast

Southern Hemisphere Megaptera novaeangliae undertake the longest migration, which reflect their exposure to lipophilic contaminants. To assess these changes, persistent organic pollutants were analyzed in blubber samples of humpback whales from three regions: the Antarctic Peninsula (n = 46), the Strait of Magellan, Chile (n = 22), and the Brazilian coast (n = 38). The similarity in PCB and HCB levels between individuals from feeding grounds and breeding grounds suggests contamination during feeding. The whales around the Antarctic Peninsula exhibited a predominance of tetrachlorobiphenyl PCBs. Whales feeding in the Strait of Magellan showed a slight prevalence of 5Cl biphenyls, likely due to their consumption of subantarctic krill species as well as small fishes potentially contaminated by industrial activities in Chile. The dominance of 5-6Cl congeners in whales in Brazil, may be attributed to the extreme physiological changes during fasting when whales utilize blubber reserves and metabolize lighter congeners, or transfer them to their calves.

Assessment of the presence of UV filters and UV stabilizers in stranded dolphin blubber

The presence of ultraviolet filters (UVFs) and stabilizers (UVSs) was evaluated for the first time in the common bottlenose dolphin (Tursiops truncatus). UVFs and UVSs are compounds of growing concern because their effects on the environment are not completely known. UVFs and UVSs are added to personal care products (PCPs), such as cosmetics and products related to sun care and once released to the aquatic ecosystem, marine organisms can bioaccumulate these substances.This work aimed to determine the presence of 12 UVFs and UVSs in cetacean blubber samples to assess the pollution to which these animals of the highest trophic chain levels are exposed due to human activity. Analytical determinations were carried out using a method based on microwave-assisted extraction combined with ultrahigh-performance liquid chromatography and tandem mass spectrometry detection. The developed method was successfully applied to determine the target compounds in the blubber tissues of five necropsied common bottlenose dolphins.Three of the 12 studied compounds, namely 2-ethylhexyl 2-cyano-3,3-diphenylprop-2-enoate (octocrilene, OC), 2-hydroxy-4-methoxybenzophenone (benzophenone 3, BP3) and 3-methylbutyl (E)-3-(4methoxyphenyl) prop-2-enoate (IMC), were detected in several samples. Of the identified compounds, OC was present in all the samples and at the highest concentration within the range from 52.61 ± 18.59 to 108.0 ± 11.32 ng·g−1.

Organochlorine contaminant concentrations and lipid profiles in eastern North Pacific gray whales (Eschrichtius robustus)

Organochlorine (OC) contaminant concentrations in tissues and lipid profiles in blubber are summarised for 101 gray whales (Eschrichtius robustus) from the eastern North Pacific stock. Samples were obtained from presumably healthy gray whales during a 1994 subsistence hunt in the Russian Arctic (n = 17) and also from biopsy sampling of live animals from the Washington coast (n = 38). In addition, tissues were collected from two groups of animals (1988-1991, n = 22; and 1999, n = 24) that stranded along the west coast of the USA. These whales represent a diverse group of animals with respect to lipid stores, age, gender, health and reproductive status. Information about these biological factors is necessary before contaminant concentration data can be properly interpreted. Differences in blubber lipid levels and profiles were examined among these groups of whales. Significantly higher lipid levels were found in the blubber of subsistence animals that were sampled following summer feeding in the Bering and Chukchi Seas, compared to lipid levels in the biopsied and stranded animals. Lipid class profiles from blubber of presumably healthy gray whales (i.e. from subsistence and biopsy sampling) contained primarily triglycerides and were very different from those of stranded animals that showed lipid decomposition (increased proportions of free fatty acids, cholesterol and phospholipids). Furthermore, lipid class profiles were found to be a means of estimating the quality of a blubber sample from stranded cetaceans. An examination of how biological factors (e.g. gender, reproductive status, age) contribute to interpreting the differences found in contaminant concentrations among the gray whales was also undertaken. Although not statistically significant, higher (OC) concentrations were found in males compared to females, thus suggesting the tendency of the mother to shift her contaminant burden to her calf during gestation and lactation. Results also indicated that there was no significant increase in concentrations of contaminants in the blubber with increase in length (surrogate for age). Higher concentrations of OC contaminants were found in stranded juvenile gray whales, compared to juvenile subsistence whales, and were thought to result from retention of OCs in blubber of the stranded animals as lipid stores are mobilised for energy and total lipid levels decrease, rather than from a difference in diet or feeding areas. OC concentrations in various tissues (blubber, liver, kidney, muscle, brain) were similar on a lipid weight basis, except for brain, which had lower lipid-adjusted OCs because the blood-brain barrier limits contaminant transfer.

Interpretation of southern hemisphere humpback whale diet via stable isotopes; implications of tissue-specific analysis.

Blubber and skin are commonly used tissues in stable isotope analysis for the purpose of investigating cetacean diet. Critical comparison of tissue-specific isotopic signals is, however, lacking resulting in uncertainty surrounding the representativeness and therefore utility of different tissues for accurate determination of recent foraging. This study used remotely biopsied blubber and skin tissues from southern hemisphere humpback whales for strategic comparison of δ13C and δ15N values. Samples were collected between 2008-2018 as part of long-term monitoring under the Humpback Whale Sentinel Program. Blubber tissues were lipid-extracted prior to analysis, whilst mathematical lipid-correction was performed on skin samples. Isotopic values from paired blubber and skin samples from the same individuals were compared to assess whether tissues could be used interchangeably for isotope analysis and dietary interpretation. Significant differences were observed for both δ13C and δ15N, flagging previously undocumented methodological considerations, and the need for method validation and standardisation in application of these approaches. This study therefore advances methodological aspects of cetacean dietary analysis. This is of elevated importance in the context of rapidly changing ocean ecosystems.

Variation in blubber thickness and histology metrics across the body topography of a false killer whale (Pseudorca crassidens).

Blubber is a multifunctional tissue essential to the survival of cetaceans. Histological assessment of blubber may be useful in determining odontocete nutritional state but a greater understanding of specific variation across the body is needed. We report on morphological variation of the blubber according to girth axes and sampling planes in a sub-adult male, bycaught false killer whale (Pseudorca crassidens) using metrics of blubber thickness (BT), adipocyte area (AA), and adipocyte index (AI). 48 full depth blubber samples were taken along 6 girth axes at 5 equidistant sampling points on both sides of the body. At these sampling locations BT was recorded, and AA and AI were determined for three distinct blubber layers. Linear mixed effect models were used to assess variation of the blubber across layers and body topography. BT was somewhat non-uniform across the body but was generally thicker in the dorsal region and thinner laterally. AA was greater cranially and AI was greater caudally. The middle and inner layer blubber showed significant differences dorsoventrally with larger AA and smaller AI in the ventral region of the body. Variation of the blubber metrics across the body are indicative of variable functions of the blubber within an individual. Due to the variability observed, we expect that AI of the dynamic inner layer blubber is most informative of overall body condition and that biopsy samples of the outer and middle blubber may still be useful in determining the nutritional status of live false killer whales.

Stratification of lipids, fatty acids and organochlorine contaminants in blubber of white whales and killer whales

The biopsy 2 via dart, trocar or surgery 2 is becoming the preferred protocol for sampling skin and blubber of many cetacean species, because a small sample from a healthy animal may provide better information than a larger sample collected via necropsy from an ill or emaciated animal. Furthermore, the biopsy is often the only means of obtaining samples (e.g. for threatened or endangered species). Because biopsy darts collect only a small sample of tissue 2 and blubber can be heterogeneous in structure and composition 2 it is essential to compare the results obtained from biopsies to those found by analysing full-thickness blubber samples obtained via necropsy. This manuscript compares blubber stratification in two odontocete species, white whales (Delphinapterus leucas) and killer whales (Orcinus orca). Five parameters (i.e. lipid percent and classes, contaminant concentrations and profiles, fatty acid profiles) were measured by blubber depth. Results of these comparisons strongly suggest that biopsy results must be interpreted with caution and in conjunction with results from species-specific blubber depth profiling. For example, lipid classes measured in biopsy samples of white whales and killer whales were similar to those for equivalent-depth samples obtained by necropsy. In addition, lipid-adjusted contaminant concentrations measured in dart or trocar samples adequately represented those obtained by necropsy of both species. Conversely, the lipid content in biopsy samples was lower than that found in same-depth necropsied samples due to loss of lipid during sampling. Also, because of the high level of fatty acid stratification observed, fatty acid profiles from the outer blubber layer collected via biopsy from both species are less likely than the metabolically active inner layer to be useful in determining the prey species consumed by these odontocetes. This study demonstrates, for white and killer whales, that properly interpreted results from blubber biopsies can provide valuable information about the body condition, health and life history of individual animals.

Strandings, mortality and morbidity of Indo-Pacific humpback dolphins in Hong Kong, with emphasis on the role of organochlorine contaminants

Factors related to mortality and disease in Indo-Pacific humpback dolphins (Sousa chinensis) from Hong Kong waters were investigated by detailed examination of dolphin specimens found stranded from 1995-2004. In total, 86 specimens were necropsied, but many of these were too badly decomposed to provide much information. Skin and blubber biopsies were also collected from six identified living individuals and concentrations of organochlorines (DDTs, PCBs and HCHs) were determined from blubber samples of stranded and biopsied dolphins. A large proportion of the strandings (53.2%) were young-of-the-year. The most commonly diagnosed causes of death were net entanglement and vessel collision. The pesticide DDT showed the highest concentrations and the ratio of DDT to its breakdown products (and other information) suggests that there may be a recent or nearby source of DDT into the dolphins’ ecosystem. Concentrations of both DDTs and PCBs showed a pattern of increasing with age in males. In females, they increased until sexual maturity, then decreased, and finally increased again in late life. This is consistent with a hypothesised transfer of pollutants from mother to offspring during gestation and lactation. Inter-laboratory differences and effects of decomposition of specimens are two potential biases that may significantly affect the quality of the present data. In order to resolve the potential problems associated with these issues, a long-term biopsy collection programme has recently been initiated.

Use of hormones in assessing reproductive physiology of humpback whales (Megaptera novaeangliae) from Juneau, Alaska

Humpback whales (Megaptera novaeangliae) in Southeast Alaska have been studied for over 50 years, and are largely considered a recovery success since the cessation of commercial whaling. Reproductive physiology is an important factor to consider in studying population health and can provide important insights into the drivers contributing to population abundance fluctuations. Validated assays for progesterone and testosterone were used on blubber biopsies from humpback whales (N = 33 whales, 71 samples) near Juneau, Alaska, in 2020 and 2021. Long-term sighting histories were used to confirm detected pregnancies with calf sightings the following year. Blubber samples were divided into two seasonal bins (early and late summer). Pregnant females sampled in both early and late summer of both 2020 and 2021 showed elevated progesterone concentrations compared to other reproductive states (p < 0.05). Progesterone concentrations in adult male whales (0.3 ± 0.2 ng/g) were not significantly different from lactating or resting female whales. Blubber testosterone concentrations in adult male humpback whales ranged from 0.05 to 1.1 ng/g, and mean concentrations were approximately double those of female whales in any reproductive state. Pregnancy was detected in 5 of 11 and 4 of 9 adult females in 2020 and 2021 respectively, yielding summer season pregnancy rates for sexually mature females at 0.45, and 0.44, respectively. Calving rates were 0.36 and 0.22 in 2020 and 2021, respectively, and the annual growth rate for this subpopulation was calculated at 2.6 % per annum. One female had successful pregnancies for four consecutive years. These results demonstrate the synergistic value of combining immunoreactive assays and long-term sighting histories to further knowledge of reproductive physiology in individual humpback whales, which can be expanded to assessing the health of a population or ecosystem.

Pregnancy rate and reproductive hormones in humpback whale blubber: Dominant form of progesterone differs during pregnancy

To better understand reproductive physiology of humpback whales Megaptera novaeangliae that reside in Hawai’i and Alaska, enzyme immunoassays were validated for both progesterone and testosterone in free-ranging and stranded animals (n = 185 biopsies). Concentrations were analyzed between different depths of large segments of blubber taken from skin to muscle layers of stranded female (n = 2, 1 pregnant, 1 non-pregnant) and male (n = 1) whales. Additionally, progesterone metabolites were identified between pregnant (n = 1) and non-pregnant (n = 3) females using high pressure liquid chromatography (HPLC). Progesterone concentrations were compared between juvenile (i.e., sexually immature), lactating, and pregnant females, and male whales, and pregnancy rates of sexually mature females were calculated. Based on replicate samples from ship struck animals collected at 7 depth locations, blubber containing the highest concentration of progesterone was located 1 cm below the skin for females, and the highest concentration of testosterone was in the skin layer of one male whale. HPLC of blubber samples of pregnant and non-pregnant females contain different immunoreactive progesterone metabolites, with the non-pregnant female eluate comprised of a more polar, and possibly conjugated, form of progesterone than the pregnant female. In females, concentrations of progesterone were highest in the blubber of pregnant (n = 28, 28.6 ± 6.9 ng/g), followed by lactating (n = 16, 0.9 ± 0.1 ng/g), and female juvenile (n = 5, 1.0 ± 0.2 ng/g) whales. Progesterone concentrations in male (n = 24, 0.6 ng/g ± 0.1 ng/g) tissues were the lowest all groups, and not different from lactating or juvenile females. Estimated summer season pregnancy rate among sexually mature females from the Hawai’i stock of humpback whales was 0.562 (95 % confidence interval 0.528–0.605). For lactating females, the year-round pregnancy rate was 0.243 (0.09–0.59), and varies depending on the threshold of progesterone assumed for pregnancy in the range between 3.1 and 28.5 ng/g. Our results demonstrate the synergistic value added when combining immunoreactive assays, HPLC, and long-term sighting histories to further knowledge of humpback whale reproductive physiology.

Investigating the relationship between the skin microbiome and flame retardant exposure of the endangered St. Lawrence Estuary beluga

The endangered beluga (Delphinapterus leucas) population in the St. Lawrence Estuary (SLE) in eastern Canada, the largest estuary in the world, is declining. Elevated tissue concentrations of a wide range of environmental contaminants, for example, halogenated flame retardants (HFRs) including polybrominated diphenyl ethers (PBDEs), might play a role in the non-recovery of this population. In mammals, HFRs have been reported to impair the metabolic regulation, including amino acid and fatty acid pathways. In the present study, we collected both blubber and skin swab samples from tissue biopsies from 56 adult SLE belugas and analyzed their blubber for the concentrations of a comprehensive suite of PBDEs and other HFRs. Using 16S rRNA marker and shotgun metagenomic approaches using skin swabs, we investigated, for the first time, the SLE beluga skin microbiome and the SLE water microbiome, providing valuable comparative taxonomic and functional microbiome information. We found that belugas have a unique skin microbiome that is distinct from surrounding SLE water, regardless of the beluga sex or location in the SLE. We further characterized the core microbiome of SLE beluga skin and surrounding SLE water, and identified bacterial taxa and gene functional pathways associated with the skin microbiome that correlated with beluga blubber HFR concentrations. Namely, we identified the phylum Nitrospinae and candidate phylum PAUC34f as potential taxa of interest that are associated with blubber HFR concentrations. We hypothesize that the biodegradation of HFRs within the beluga blubber and skin results in an increase in local metabolite concentrations that leads to the proliferation of Nitrospinae and PAUC32f. This work demonstrates the utility of studying the core microbiome of the SLE beluga skin using a swab method that could be adapted to field sampling. Further studies of the temporal effects of contaminant exposure on SLE beluga skin and SLE water microbiomes is warranted for potentially better monitoring and protecting this marine mammal which is at risk.

Temporal trends of persistent organic pollutant contamination in Franciscana dolphins from the Southwestern Atlantic

Persistent organic pollutants (POPs) were analyzed in 136 blubber samples of Franciscana dolphins from Brazil (Pontoporia blainvillei), which is the most threatened dolphin in the Southwestern Atlantic. The dolphins were caught by the fishery fleet and collected from 2000 to 2018 in three regions of São Paulo state: northern São Paulo (SPN), central São Paulo (SPC), and southern São Paulo (SPS). The POPs analyzed in this study were polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDTs), Mirex, hexachlorobenzene (HCB), chlordane compounds (CHLs), hexachlorocyclohexane isomers (HCHs), and polybrominated diphenyl ethers (PBDEs). The concentrations ranged from 36 to 7200 ng g−1 lipid weight (lw) and 113–42200 ng g−1 lw for predominant compounds DDTs and PCBs, respectively. Similar profiles of PCB congeners were observed with a predominance of hexachlorinated compounds, representing approximately 50% of the total PCB amount; the highest PCB concentrations were observed from Baixada Santista (SPC) proximate to a highly urbanized and industrial coastal area. Significant differences were observed between the sexes and maturity of dolphins, mainly for PCBs, DDTs, and Mirex. In general, POPs other than HCB in Franciscana dolphins showed downward temporal trends, matching the regulatory periods for restricting and/or banning these compounds. Although POP concentrations are declining, PCB levels remain high in small dolphins, suggesting adverse health effects on Franciscanas. As organic contaminants are one of the numerous threats Franciscanas have been vulnerable to along the Brazilian coast, we recommend monitoring POPs levels every five years to check for declining (or stabilizing) trends.

Blubber and serum cortisol concentrations as indicators of the stress response and overall health status in striped dolphins

The impacts of environmental changes and anthropogenic threats in marine mammals are a growing concern for their conservation. In recent years, efforts have been directed to understand how marine mammals cope with stressors and to assess and validate stress biomarkers, mainly levels of glucocorticoid hormones (e.g. cortisol) in certain body tissues. The aims of this study were to assess the impact of different causes of stranding (chronically affected and bycaught striped dolphins) on cortisol concentrations in serum and in blubber; and to evaluate the association between cortisol levels in these tissues. Blubber and blood samples were collected from striped dolphins (n = 42) stranded on the Mediterranean coast between 2012 and 2018. Cortisol concentrations were measured by using enzyme immunoassay. A high correlation was found between circulating and blubber cortisol concentrations (R2 = 0.85, p < 0.01). Necropsies and pathological studies concluded that a third of the dolphins were bycaught in fishing nets and released by fishermen (Bycaught animals group), while the other two thirds were euthanized, or died, due to a disease or chronic condition (e.g. calves separated from the mother or animals infected with dolphin morbillivirus or Brucella ceti) that impeded survival (Chronically affected animals group). Cortisol concentrations (mean ± SD) were six times higher in chronically affected animals (35.3 ± 23 ng cortisol/g blubber and 6.63 ± 3.22 μg cortisol/dl serum) compared to those bycaught in fishing nets (6.2 ± 4.3 ng cortisol/g blubber and 1.15 ± 1.51 μg cortisol/dl serum). Results suggests that serum and blubber cortisol concentrations can contribute in inferring the overall health and welfare of free-ranging cetaceans. However, further research is required to understand better the kinetics of blubber cortisol incorporation and removal, the factors involved in these processes, and the local conversion of cortisol in the blubber.